Contributions
Abstract: PB2340
Type: Publication Only
Background
Thienopyridines are a family of compounds well known for their anti-platelet activity, and are commonly used to prevent thrombotic events. However, attention has recently turned to their anti-cancer properties. Mechanistic investigations have explored their mode of action, with PLCγ, Jak2 and also Lck suggested as direct targets, while mitotic spindle inhibition via micro-tubule de-stabilisation has also been demonstrated. Mature B-cell malignancies are infamously difficult to treat. Traditionally, treatment approaches have included Microtubule Target Agents (MTA) such as vincristine and vinblastine that bind to tubulin and disrupt microtubule dynamics. MTA have been hugely successful and are among the most important anti-cancer drugs. However, neurotoxicity, neutropenia and the development of drug resistance can be limiting factors. A greater understanding of the mechanisms of action of MTAs will enhance their use and effectiveness and aid in the identification of newer compounds with this action.
Aims
The aim of this study was to assess the effects of six novel thienopyridine derivatives synthesized by our group on B-cell apoptosis, cell cycle arrest and mitotic disruption with the intention of identifying a new family of compound for use in the treatment of mature B-cell neoplasms
Methods
The Burkitt lymphoma mature B-cell line, Daudi, was used in this study. Cells were seeded at a standard density dependent on the downstream assay to be performed and treated with varying concentrations (1-100µM) of thienopyridine derivative (DJ0081, DJ0199, DJ0206, DJ0209, DJ0014 and DJ0021) for varying culture times. Cellular biochemical activity was assessed using the MTS assay, while apoptosis and necrosis were observed using the Annexin V/Propidium Iodide flow cytometry assay. Cell cycle arrest was determined by flow cytometric Propidium Iodide cell cycle assay, and alpha-tubulin expression was visualised using confocal microscopy
Results
Five of the novel compounds demonstrated a significant reduction in biochemical activity after 48h and 72h of treatment at 10µM with three of the compounds having affects at 1µM, indicating cytostasis/cell death. As expected, compound DJ0021 (synthesised as the control compound) did not significantly affect cell viability. At these same concentrations and time points, significant apoptosis was induced, with very few necrotic events observed by flow cytometry. Cell cycle analysis demonstrated a significant G2 arrest and tetraploidy following just 24h treatment with 10µM, and more marked G2 arrest at 48 and 72h treatment, while microtubule formation was found to be disrupted by all five compounds.
Conclusion
The study demonstrates the anti-mitotic activity of these thienopyridine derivatives and their potential for use in the treatment of mature B-cell malignancies,
Session topic: 19. Non-Hodgkin lymphoma Biology & Translational Research
Keyword(s): Apoptosis, B cell, Cell cycle
Abstract: PB2340
Type: Publication Only
Background
Thienopyridines are a family of compounds well known for their anti-platelet activity, and are commonly used to prevent thrombotic events. However, attention has recently turned to their anti-cancer properties. Mechanistic investigations have explored their mode of action, with PLCγ, Jak2 and also Lck suggested as direct targets, while mitotic spindle inhibition via micro-tubule de-stabilisation has also been demonstrated. Mature B-cell malignancies are infamously difficult to treat. Traditionally, treatment approaches have included Microtubule Target Agents (MTA) such as vincristine and vinblastine that bind to tubulin and disrupt microtubule dynamics. MTA have been hugely successful and are among the most important anti-cancer drugs. However, neurotoxicity, neutropenia and the development of drug resistance can be limiting factors. A greater understanding of the mechanisms of action of MTAs will enhance their use and effectiveness and aid in the identification of newer compounds with this action.
Aims
The aim of this study was to assess the effects of six novel thienopyridine derivatives synthesized by our group on B-cell apoptosis, cell cycle arrest and mitotic disruption with the intention of identifying a new family of compound for use in the treatment of mature B-cell neoplasms
Methods
The Burkitt lymphoma mature B-cell line, Daudi, was used in this study. Cells were seeded at a standard density dependent on the downstream assay to be performed and treated with varying concentrations (1-100µM) of thienopyridine derivative (DJ0081, DJ0199, DJ0206, DJ0209, DJ0014 and DJ0021) for varying culture times. Cellular biochemical activity was assessed using the MTS assay, while apoptosis and necrosis were observed using the Annexin V/Propidium Iodide flow cytometry assay. Cell cycle arrest was determined by flow cytometric Propidium Iodide cell cycle assay, and alpha-tubulin expression was visualised using confocal microscopy
Results
Five of the novel compounds demonstrated a significant reduction in biochemical activity after 48h and 72h of treatment at 10µM with three of the compounds having affects at 1µM, indicating cytostasis/cell death. As expected, compound DJ0021 (synthesised as the control compound) did not significantly affect cell viability. At these same concentrations and time points, significant apoptosis was induced, with very few necrotic events observed by flow cytometry. Cell cycle analysis demonstrated a significant G2 arrest and tetraploidy following just 24h treatment with 10µM, and more marked G2 arrest at 48 and 72h treatment, while microtubule formation was found to be disrupted by all five compounds.
Conclusion
The study demonstrates the anti-mitotic activity of these thienopyridine derivatives and their potential for use in the treatment of mature B-cell malignancies,
Session topic: 19. Non-Hodgkin lymphoma Biology & Translational Research
Keyword(s): Apoptosis, B cell, Cell cycle